<p>Theabrownin Induces Apoptosis and Tumor Inhibition of Hepatocellular Carcinoma Huh7 Cells Through ASK1-JNK-c-Jun Pathway</p>

Abstract

Purpose Theabrownin (TB), a main pigment and bioactive component of tea, has been shown anti-tumor activities against carcinomas, but its effects on hepatocellular carcinoma (HCC) remain unclear. Methods Hepatocellular carcinoma Huh7 cells were used for analyses. Cell viability assay was performed to determine TB′s anti-proliferative effect, and flow cytometry with annexin V-FITC/PI double staining and DAPI staining were performed to determine its pro-apoptotic effect. Real-time PCR and Western blot assays were conducted to detect the molecular actions of TB. And a xenograft model of zebrafishes was established to evaluate the in vivo effect of TB. SP600125 (JNK inhibitor) was in vivo and in vitro used to verify the regulatory role of the JNK signaling pathway in the anti-hepatic carcinoma mechanism of TB. Results TB exerted significant anti-proliferative and pro-apoptotic effects on Huh7 cells in a dose-dependent manner. The molecular data showed that TB up-regulated the gene expressions of NOXA, PUMA, P21, Bax, and Bim and up-regulated the protein expressions of ASK-1, Bax, phosphorylated JNK, and phosphorylated c-Jun with down-regulation of Bcl-2. The in vivo data showed that TB exerted significant tumor-inhibitory effect which was even stronger than that of cis-platinum. Furthermore, the JNK inhibitor significantly weakened TB′s effects both in vivo and in vitro and blocked the related molecular pathway. Conclusion TB exerts anti-proliferative, pro-apoptotic, and tumor-inhibitory effects on Huh7 cells through activation of the JNK signaling pathway. For the first time, this study provides new evidence of anti-HCC effects and mechanism of TB.

Green tea-derived theabrownin induces cellular senescence and apoptosis of hepatocellular carcinoma through p53 signaling activation and bypassed JNK signaling suppression

“…Alike most natural products, TBʹs in vitro effects on cancer cells are milder than chemotherapeutics, with higher IC 50 s. However, this study demonstrated that TB had considerable efficacy against HCC cell mass in xenograft zebrafish and the in vivo efficacy was even better than Cis-platin. Our finding was in consistent with previous reports about the in vivo anti-cancer study of TB [ 37 , 39 ], which suggested that TB might not only induce cancer cell apoptosis in a direct way, but also suppress in vivo cancer cell mass in an indirect way (e.g., anti-angiogenesis and immunoregulation) [ 63 – 65 ]. Further studies are needed to explore the in vivo mechanism of TB.…”

“…Our previous studies have shown that TB induced apoptosis on p53-mut HCC cells via activation of JNK signaling [ 39 ]. Thus, JNK-siRNA was applied to verify the involvement of JNK in the action mechanism of TB.…”

“…7 ). In our previous study, TB induced apoptosis in p53-mut Huh7 cells via activation of JNK signaling pathway, indicating that JNK might also participate into the action mechanism of TB on HCC cells [ 39 ]. Correspondingly, we revealed that JNK was a bypass regulator involved in the mechanism of TB (Fig.…”

Sorafenib-induced Aquaporin-3 Downregulation is Coupled With Proliferation Inhibition and Increased Apoptosis in Hepatocellular Carcinoma Cells

“…Cell culture, reagents, and antibodies Human HCC cell lines Huh7, HepG2 and SK-Hep1 obtained from The Chinese Academy of Sciences in Shanghai-China were used. Cells were cultured in Dulbecco's Modi ed Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (Gibco, Shanghai, China) and 1% penicillin/streptomycin and maintained in an incubator with a humidi ed atmosphere of 5% CO 2 at 37°C 30,31 . Sorafenib and UO126 (Sigma-Aldrich, St. Louis, Missouri, USA) were purchased and dissolved in 100% DMSO to prepare a 10mM stock solution, which was then diluted with DMEM to the desired concentration, with a nal concentration of 0.1% DMSO, recommended for in vitro studies.…”

Xenograft Zebrafish Models for the Development of Novel Anti-Hepatocellular Carcinoma Molecules

“…Its anticancer potential has been proposed [ 65 ]; however, the effectiveness in HCC needs investigation. In cultured HCC cell lines, HepG2 (well differentiated HCC cells [ 55 ]), HuH7 and SKHep1 (cell line of endothelial origin, not of HCC origin [ 56 ]), Xu et al [ 43 ] showed that TB (200 μg/μL) potently reduced cell viability, as evaluated by MTT test. This was due to the activation of the JNK signaling pathway that resulted in an anti-proliferative and pro-apoptotic effect.…”